Antibody recognizing human 25-hydroxyvitamin D3-1a-hydroxylase

Abstract

The present invention relates to a polypeptide having 25-hydroxyvitamin D 3 -1α-hydroxylase activity, being useful for the prevention, diagnosis and therapeutic treatment of adult diseases such as osteoporosis induced by the decrease of active type vitamin D 3 and catalyzing the final stage of vitamin D 3 activation; and the gene encoding the polypeptide. In accordance with the present invention, the following can be provided; a polypeptide having 25-hydroxyvitamin D 3 -1α-hydroxylase activity, DNA encoding the polypeptide, a recombinant DNA prepared by inserting the DNA in a vector, a transformant carrying the recombinant DNA, a method for preparing 25-hydroxyvitamin D 3 -1α-hydroxylase by using the transformant, a method for preparing 1α, 25-dihydroxyvitamin D 3 comprising using the polypeptide having 25-hydroxyvitamin D 3 -1α-hydroxylase activity, and an antibody recognizing the polypeptide.

What is claimed is:  
     
         1 . A polypeptide comprising an amino acid sequence selected from the amino acid sequences represented by SEQ ID NOS. 1 and 2, or 
 a polypeptide comprising an amino acid sequence in which amino acids residues are deleted, substituted or added in the amino acid sequence of said polypeptide, and having 25-hydroxyvitamin D 3 -1α-hydroxylase activity.    
     
     
         2 . A DNA encoding a polypeptide according to  claim 1  or 
 a DNA which hybridizes with said DNA under stringent conditions.  
 
     
     
         3 . The DNA according to  claim 2 , wherein the DNA is DNA comprising a nucleotide sequence selected from nucleotide sequences represented by SEQ ID NOS. 3 and 4.  
     
     
         4 . A recombinant DNA prepared by inserting the DNA according to  claim 2  or  3  into a vector.  
     
     
         5 . A transformant carrying a recombinant DNA according to  claim 4 .  
     
     
         6 . A method for producing 25-hydroxyvitamin D 3 -1α-hydroxylase, comprising: 
 cultivating the transformant according to  claim 5  in a medium to produce 25-hydroxyvitamin D 3 - 1α-hydroxylase in the culture; and  
 recovering said 25-hydroxyvitamin D 3 -1α-hydroxylase from the resulting culture.  
 
     
     
         7 . A method for producing 1α, 25-dihydroxyvitamin D 3 , comprising: 
 putting the polypeptide according to  claim 1  and 25-hydroxyvitamin D 3  in an aqueous medium to produce 1α, 25-dihydroxyvitamin D 3  in the aqueous medium; and  
 recovering said 1α, 25-dihydroxyvitamin D 3  from the aqueous medium.  
 
     
     
         8 . An antibody recognizing the polypeptide according to  claim 1 .  
     
     
         9 . A method for immunologically detecting a polypeptide having 25-hydroxyvitamin D 3 -1α-hydroxylase activity, using the antibody according to  claim 8 .  
     
     
         10 . An immuno-histological staining method comprising using an antibody according to  claim 8 .  
     
     
         11 . An immuno-histological staining agent containing an antibody according to claim  8 .